| Objective: To demonstrate that SLE T cells primary function disordor is relative to abnormal biochemical pathways, and to furthermore search the pathogenesis of SLE. Methods: Human PBMC were isolated by standard Ficoll-Hypaque gradient centrifugation from heparinized peripheral blood from SLE patients and control individuals, T lymphocytes were isolated and enriched from PBMC by Nylon immunoafflnity columns, cytofluorographic analysis of enriched T lymphocyte preparations with FITC-sheep anti mouse IgG and anti-CD3- c demonstrated that over 90% possessed the CD3 membrane complex that defines T lymphocytes. The UCHTI anti-CD3 mAb elicits Ca response in T cells with a second cross-linking sheep anti-mouse Ab lgG by ACAS, Base line., peak and 5-mm (plateau) [Ca]i responses were recorded. The Percentage of CD3~ cells among T cells were analyzed by flow cytometry. we monitored [Caji responses in SLE and normal T cells after addition of 1 ii M thapsigargin, a inhibitor of the sarcoplasmic and endoplasmic Ca--ATPase, Anti-CD3-induced IICa1i response of SLE and normal T cells was monitored in presence of 5mM EGTA, the extracellular Ca~~ chelator. InsP3 levels were measured in T cells from SLE patients and control by using a sensitive radioassay. Fresh T lymphocytes were prepared as above, SLE and control T cells were activated in vitro by monoclonal anti-CD3- r antibody cross-linking sheep anti-mouse antibody IgG and recombinant IL-I ci (rIL- 1 ci ) for 24 hours, T cell activation was verified by JL-2 mRNA. IL-2R- a mRNA and IL-2R- 13 mRNA expression. Total PKA activity of I cells was quantified by liquid scintillation spectrometry, control T cells cultured for 24 hours in medium Supplemented with 10% active SLE Sera or increasing concentrations of IFN- a and total PKA activity was quantified as above. Results: 1. The base line recordings of [CaJi response from SLE patients were similar to that from normal control (P<0.74). 2. Peak [Ca~t]i response wre significantly higher in the group of patients with SLF than that in the control group (p<0.004),Similarly, the 5-mm response was significantly higher in SLF group than that in the normal control group (p |
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